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US 2023/0416723 A1Published application
Methods of oligonucleotide-based affinity chromatography
AssigneeDonaldson Company, Inc.
InventorsJinxiang ZHOU, Graham TEMPLES
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Abstract
Disclosed are methods for purifying polynucleotides from a feed solution using oligonucleotide affinity columns at flowrates between 0.5 CV/min to 1000 CV/min. The methods can include several steps including loading a mixture containing the target polynucleotide onto a chromatography media, e.g., a column, that carries a macroporous support with an oligonucleotide affinity ligand bonded to a surface. The affinity ligand can hybridize the targeted polynucleotide and allow for separation and purification of the target.
Claims20
A method for purifying a polynucleotide comprising:
- loading a feed solution comprising the polynucleotide onto a chromatographic media, the chromatographic media comprising a macroporous support having a specific surface area of from about 1 m2/mL to about 20 m2/mL, the macroporous support including an oligonucleotide affinity ligand bonded to a surface of the macroporous support and in fluid communication with the feed solution, the oligonucleotide affinity ligand comprising a nucleotide sequence that is complementary to a nucleotide sequence of the polynucleotide, the chromatographic media exhibiting a dynamic binding capacity of from about 0.2 mg polynucleotide/mL to about 15 mg polynucleotide/mL;
- flowing the feed solution through the chromatographic media, wherein upon the flowing, the polynucleotide is retained on the macroporous support via hybridization with the affinity ligand and impurities of the feed solution pass through the chromatographic media;
- and
- collecting the polynucleotide following separation of the polynucleotide from the macroporous support.
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Dependencies are inferred from the claim text.